The release of the ICH M12 Guideline on Drug Interaction Studies has reignited discussions around assay validation requirements for in vitro assays such as plasma protein-binding studies. Even though the ICH M12 does not directly reference the ICH M10 Guideline on Bioanalytical Method Validation and Sample Analysis, its release prompted further discussions on assay validation requirements for these studies during the 17th European Bioanalysis Forum Open Symposium held in Barcelona, Spain, from 19 to 21 November 2024, where we advocated for a Context-of-Use driven approach over rigid adherence to ICH M10 standards. Context-of-Use driven validation ensures assays are tailored to the specific scientific and regulatory needs, optimizing resource allocation and innovation in drug development. This short opinion paper explores the potential and undesired implications of ICH M12 on bioanalytical validation practices, highlights the distinction between exploratory assays and assays having a clinical impact, and underscores the necessity for tailored validation strategies.

This manuscript reports back from the discussion in the European Bioanalysis Forum community on the challenges observed when implementing Good Clinical Practices in the bioanalytical laboratory. It is not intended to challenge any regulatory requirements but to open a discussion on where the bioanalytical community sees ambiguities on implementing Good Clinical Practices or areas where expectations are either felt not being owned by Bioanalysis or where Good Clinical Practices requirements are at risk of getting contaminated with requirements originating from Good Laboratory Practices. In addition to this, the discussions focused on three additional main challenges: the informed consent withdrawal, expedited reporting of unexpected results and the risk-based approach to quality management, The European Bioanalysis Forum community is continuing discussions, but already this manuscript should help to appreciate the challenges and to try and resolve them, involving all stakeholders.

The ICH M10 guideline on bioanalytical method validation and sample analysis is being adopted since 2023. However, and inevitably, some paragraphs or requirements remain ambiguous and are open for different interpretations. In support of a harmonized interpretation by the industry and health authorities, the European Bioanalysis Forum organized a workshop on 14 November 2023 in Barcelona, Spain, to discuss unclear and/or ambiguous paragraphs which were identified by the European Bioanalysis Forum community and delegates of the workshop prior to the workshop. This manuscript reports back from the workshop with recommendations and aims at continuing an open scientific discussion within the industry and with regulators in support of a science-driven guideline for the bioanalytical community and in line with the ICH mission - that is, achieve greater harmonization worldwide to ensure that safe, effective and high-quality medicines are developed and registered in the most resource-efficient manner.

In this report, the European Bioanalysis Forum shares the proposals for harmonized implementation of the ICH M10 guideline on bioanalytical method validation and study sample analysis from the ICH M10 workshop. The focus of the discussions was to understand new, changed or still ambiguous regulatory expectations in the guideline, as identified in feedback from the pre-workshop surveys or during the workshop. The proposals from the workshop aim at stimulating and helping a harmonized implementation of the guideline, and using our community as a sounding board during and after implementation to highlight areas of misalignment and to create a platform for continued sharing with the regulatory authorities in an effort to contribute to industry and regulators developing similar interpretations on guideline expectations.

Quit Victoria (a program of Cancer Council Victoria) developed, in partnership with the Heart Foundation and VicHealth, a multichannel advertising campaign to raise awareness of platelet aggregation ('sticky blood') caused by smoking. The primary audiences were English-speaking people who smoke (aged 29-54 years) and Chinese-Victorians who smoke. The secondary audience was general practitioners, cardiac specialists, and nurses.

Once released, the ICH M10 Guideline on bioanalytical method validation will become one of the most important milestones in the history of regulated bioanalysis, closing a chapter on intense discussions among the industry and health authorities started in Crystal City in 2001. In this manuscript, the European Bioanalysis Forum community reports back on their feedback on the ICH M10 draft guideline gathered during the public consultation period. The comments given are intended to contribute to a guideline that combines several decades of experience and current scientific vision. They should provide future generations of bioanalytical scientist a regulatory framework so their bioanalytical work can contribute to safe, effective and high-quality medicines, which can be developed and registered in the most resource-efficient manner.

Purpose of study The United States is experiencing an opioid overdose epidemic. Since 2010, overdose deaths have increased by more than 200% due in part to the introduction of illicit fentanyl which can cause acute respiratory failure. The overdose rescue drug naloxone has saved thousands of lives. Yet illicit opioid users often use alone, which increases risk of fatal overdose by decreasing opportunity for intervention. One potential solution (developed by our research team) is a contactless self-monitoring smartphone app which can detect respiratory depression and call naloxone-equipped friends, family or emergency medical services. A key challenge of self-monitoring is minimizing false alarms; to address this, we sought to measure opioid users’ ability to interact with a smartphone after an opioid injection (and thus their ability to terminate a false alarm). We hypothesized that when users were impaired but not in acute distress, they would be able to interact with a smartphone. Methods used We conducted a pilot study at the InSite Supervised Injection Facility in Vancouver, Canada. Opioid users were consented by InSite staff and taught how to turn off a smartphone alarm. Participants were monitored for 30 s prior to injection, and a 2 min smartphone countdown alarm was set upon injection completion. Monitoring continued for 30 s post-alarm. Then researchers explained the concept of a smartphone-based self-monitor which could call for help if an overdose was detected. Participants were asked if they would use such a device and how many ‘false alarms’ they could reasonably tolerate. Summary of results Of 42 participants who successfully injected, 83% turned off the alarm within 30 s, 7% tried to turn it off but had interface challenges, and 9% made no effort. None of the participants experienced an overdose. Of 50 participants asked if they would use such an app, 72% responded ‘yes’, 12% ‘maybe’, and 6% ‘no’ (the remaining 10% declined to answer). Conclusions Our pilot study tested opioid users’ ability to interact with a smartphone while impaired. Results suggest that >90% of opioid users are able to interact with a smartphone in the minutes following self-injection. We conclude it would be reasonable to expect users to turn off a smartphone alarm if they were not experiencing life-threatening respiratory depression.

Aim: Microsampling in preclinical pharmacokinetics (PK) studies is currently widely adopted across the pharmaceutical industry. Materials & methods: The European Bioanalysis Forum liquid microsampling consortium member companies assessed the accuracy and precision of handheld pipettes and microcapillaries at volumes of less than 10 μl. The following key factors on pipetting performance were also evaluated: Pipette type (positive displacement, air displacement and microcapillary), experience of user and the liquid type. Water was selected as a best-case scenario for accuracy and precision determination and blood plasma as a 'real world' bioanalysis sample type. Conclusion: Accuracy and precision on the pipetted volume decreased at lower volumes and experienced laboratory technicians performed better compared with the infrequent users. With respect to the pipetting devices used, microcapillaries showed better or equivalent accuracy and precision compared with handheld pipettes across the volume range 1-8 μl independent of the matrix used.

The European Bioanalysis Forum is providing a proposal on experiments to be conducted if processed samples are to be stored for a period of time, either after an initial injection or if processed samples are to be stored for a period of time prior to their first injection. Using survey information and extensive discussion an insight was gained as to whether the current practices employed during method validation were conducted to scientifically validate actual sample handling conditions or to simply meet a perceived regulatory expectation. The goal of this report is to recommend the adoption of scientifically justified validation experiments that exactly reproduce and processed test sample handling and storage procedures and to introduce some uniformity across the industry.

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BioanalysisVol. 10, No. 16 White PaperFree AccessToward decision-based acceptance criteria for Bioanalytical Method Validation: a proposal for discussion from the European Bioanalysis ForumPhilip Timmerman, Michaela Golob, Joanne Goodman, Magnus Knutsson, Robert Nelson, Marianne Scheel Fjording & Steve WhitePhilip Timmerman*Author for correspondence: Tel.: +32 479 91 01 32; E-mail Address: chair@e-b-f.eu European Bioanalysis Forum, Havenlaan 86c b204, 1000 Brussels, Belgium, Michaela Golob Nuvisan, Grafing, Germany, Joanne Goodman MedImmune, Cambridge, UK, Magnus Knutsson Ferring, Copenhagen, Denmark, Robert Nelson Novimmune, Geneva, Switzerland, Marianne Scheel Fjording Novo Nordisk, Måløv, Denmark & Steve White GlaxoSmithKline, Ware, UKPublished Online:21 Aug 2018https://doi.org/10.4155/bio-2018-0131AboutSectionsPDF/EPUB ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareShare onFacebookTwitterLinkedInReddit Validation of bioanalytical assays has been widely discussed over the last decades. Since the early 1990s, industry and regulators have been working on a consensus model building on the conclusions from the conference on Analytical Methods Validation: Bioavailability, Bioequivalence and Pharmacokinetic studies [1]. This first international conference was held in December 1990 in Crystal City, Arlington, USA and was sponsored by the American Association of Pharmaceutical Scientists (AAPS), US FDA, Federation Internationale Pharmaceutique, Health Protection Branch (Canada) and Association of Official Analytical Chemists. The conference has been referred to as the Crystal City I (CC-I) conference, and became the template for subsequent Crystal City (CC) conferences held between 2000 and 2015 [2–5]. All current regional or international regulations related to validation of bioanalytical assays and industry publications on the subject were based on the principles and practices discussed and agreed at the CC-I conference. Although, Health Canada formally referred to the CC-I conference report for bioequivalence studies in 1996 [6], it was only in 2001 that the FDA actually issued a formal guidance, which has been recently updated [7]. This 2001 guidance was based largely on the discussions from the CC-II meeting, which refined the conclusions from 1990 after a decade of experience with the CC-I principles. During that decade, important progress was made in various areas of analytical chemistry. Subsequent CC meetings and other international meetings hosted by, for example, the European Bioanalysis Forum (EBF) (together with European Federation for Pharmaceutical Sciences (EUFEPS) in 2010) or the Japan Bioanalysis Forum [8] to comment and provide input into upcoming regulations in Europe and Japan, brought together the industry experts and regulators to refine the practices or agree (but sometimes also disagree) to add or modify regulatory requirements based on observed flaws in methods or enhancements in technology. Some of the added requirements were inspired by industry sharing expertise at meetings or in peer-reviewed literature, others were requested by the regulators as part of their observations during regular audits and/or inspections at the time of filings.Globalization of the Bioanalytical GuidelinesDuring the last decade, the regulated bioanalytical community truly globalized. We have also observed Bioanalytical Method Validation Guidelines emerging in many regions of the world. Most of these guidelines were based upon the aforementioned 2001 FDA guidance and included many of the conclusions from the different CC meetings. And although all guidelines were built on the same principles, some small differences from one guideline to another created a huge problem for the global bioanalytical community on how to comply with all requirements in an efficient way.Hence, since 2010, and aligned with guidelines being issued in regions outside of the USA, the industry intensified their desire on harmonization of bioanalytical guidance. At first, and as can be read from the Open Letter to the Regulators in 2011, the request for harmonization referred to harmonized interpretation of the guidance principles by the industry and the regulators [9]. Going forward, the industry united under the umbrella of the Global Bioanalysis Consortium, which in turn reported back on their discussion by issuing a number of high quality publications summarizing industry's current thinking on best practices for Bioanalytical Method Validation [10]. From there, industry's focus gradually shifted to a request for harmonization of the different regional guidelines and guidance involving the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH), a proposal that was also made by the regulators, leading to the current development of the ICH M10 guideline [11].Adding more details in the guidelinesAs mentioned in the introduction of this manuscript, since the early 1990s of the 20th century, all discussions and practices built on the principles and paradigms of the CC-I paper, discussing the criteria for chemical assays on the one hand and ligand-binding assays (LBAs) on the other hand. Already in 2001, industry and regulators considered the progress in chemical assays and included more assay-specific requirements for the booming LC–MS area, dealing with matrix effects being the most prominent assay challenge. Also, the 2001 FDA guidance was far from complete or even satisfactory for the LBA community as exemplified by the CC-II follow-up meeting and subsequent meeting report on the best practices for these assay formats [12,13]. The DeSilva publication certainly added more details for LBA assays and became the foundation for LBA assay validations for the next decade [13].New modalities & technologiesOf course, from 2001 to date, technology progress did not stop and it will not stop tomorrow. Equally important, drug R&D portfolios continue to branch into many different areas and new modalities, requiring the bioanalytical scientist to reposition existing technology used in the laboratory or to integrate completely new analytical platforms. For one, this leads to a tremendous refinement of existing technology, both for chemical and LBAs with respect to the sensitivity, specificity and selectively. But also, it requires the bioanalytical scientists to combine assay formats or introduce new techniques into their (regulatory) bioanalytical toolbox.In June 2011, the EBF hosted a Focus Meeting discussing the progress in technology allowing chemical assays to be used for a broader array of compounds historically only analyzed by binding assays [14]. One of the desires expressed at that meeting was not to simply 'copy/paste' acceptance technology-based acceptance criteria when analyzing peptides and proteins with MS-based assays, but to reflect if a cutting-edge protein analysis using LC–MS and the decisions taken with the data really need more stringent acceptance criteria than if the same molecule would have been analyzed using the conventional LBA. These were early days for peptide and protein analysis using LC–MS. During the years following the 2011 EBF Focus Meeting, industry experienced an exponential increase in technological opportunities to analyze larger molecules using MS detection and quantification. A steep learning curve, which is still continuing.Technology-based versus decision-based acceptance criteriaIn June 2017, the EBF hosted a second Focus Workshop on the theme in support of developing Bioanalytical Strategies for Large Molecules in Modern Drug Development [15]. In the meantime, many bioanalytical laboratories were developing and validating a broad array of assays for the quantification of peptides and (large) proteins using immunocapture and/or direct protein digest LC–MS(/MS) methods replacing or complementing the traditional LBA. Additionally, it has increasingly become unclear which acceptance criteria to apply, irrespective of a comprehensive AAPS paper published in 2015 [16]. During the workshop discussions, the idea developed that industry should take a step back and reflect on one of the starting principles of the Bioanalytical Method Validation: should technology drive the acceptance criteria of bioanalytical assays? During the panel discussions at the workshop, the delegates discussed if the current acceptance criteria for bioanalytical assays, and the difference when one technology is used versus another, should not be appraised to come to harmonized criteria, independent from the technology, but driven by the decision taken with the data, that is, a pharmacokinetic (PK) and/or toxicokinetic assessment. What would be the appropriate criteria be if they were not driven by what we can achieve, but by what is needed to take decisions in support of patient safety and efficacy of (new) drug products?Since the 1990 CC-I paper [1], the industry and regulators based the discussions on which acceptance criteria to apply on the technology used: chromatographic assays were held to accuracy/precision within 15% (20% at LLOQ) and binding assays at accuracy/precision within 20% (25% at LLOQ/ULOQ), irrespective of the decisions taken on the data. As part of these discussions, and not unimportant to mention, we may have lost the fact that prior to 2001 the agreed accuracy/precision was ±20% and ±25% for chromatographic/chemical assays and LBAs, respectively.The proposal made at the 2016 EBF Focus Workshop was followed by a lively discussion on redefining acceptance criteria for Bioanalytical Method Validation and basing them on the decisions taken on the data. In this way, the proposal can contribute to the harmonization efforts in the bioanalytical community: defining one harmonized level of acceptance criteria for assays used to make PK and safety decisions. A number of arguments were brought forward in support of this proposal. First, it provides an answer to a discussion, which keeps the industry busy ever since CC-II. Shortly after that meeting in 2000, where industry and regulators confirmed that they were comfortable with ±20 or 25% for chemical/chromatographic and LBA assays, respectively, the decision was taken to change acceptance criteria from 20 to 15% and 25 to 20% for the chemical/chromatographic and LBA assays, respectively. However, these tighter criteria were still technology-based and did not consider that we actually make identical PK or safety claims on these chromatography or LBA data even though they inherently may differ in accuracy and precision. Also, the accuracy and precision criteria themselves may even be irrelevant in the context of the downstream decision made on these data. Hence, we often heard the question: why should the acceptance criteria be different just because a different analytical platform is utilized?Second, harmonized decision-based acceptance criteria can provide an acceptable answer to one of the key questions from the EBF Workshop 'Which criteria to use in so-called 'hybrid assays' or immunocapture and/or protein digest LC–MS(/MS) methods?'. Are these 'hybrid assays' chromatographic assays or LBAs? And, even more challenging, why would scientists develop a cutting edge 'hybrid assay' if they will be held to more stringent chromatography acceptance criteria for data supporting exactly the same PK or safety claim as an LBA for that same compound?A third and maybe most important argument is related to the fact that new technologies and new modalities will continue to enter our world. In the coming years, other technologies will be required to provide the PK/toxicokinetics (TK) answers for regulatory review and labeling. How will we be managing or regulating the use of these new technologies? As an example, which category should be utilized for flow cytometry, qPCR, positron emission tomography (PET) imaging or the variety of new MS-based applications? Which criteria should apply when these technologies are used to support PK/safety claims similar to conventional LC–MS/MS or LBAs?As a bioanalytical community, we are aware of the game-changing impact of the proposal made in this manuscript and realize it requires intense discussions and reflections. This is also not a proposal to bring acceptance criteria for chromatography-based assays to ±20% or for LBAs to ±15%. Instead, we are making a plea to define and agree on harmonized criteria, which can support the decision made on dosing, PK and safety from the bioanalytical data. As a consequence, this discussion needs active involvement and input from the end users of these concentration data. Indeed, as the proposal is about applying decision-based acceptance criteria instead of technology-based criteria, input from the stakeholders about making these decisions is crucial. We should not start from a blank page either; we have almost three decades of experience with the current level of acceptance criteria. At the same time we should not forget that from the inception of the 4-6-XX paradigm, experts were already challenging whether this is the best way to accept/reject bioanalytical data for PK [17].The above proposal may also help progress the intense discussion on which criteria to use for biomarker assays. Since biomarker assays are pharmacodynamic assays, they should not automatically be held to PK acceptance criteria. It may help to develop pharmacodynamic acceptance criteria, which likely are individual per biomarker entity. We would like to refer to an earlier EBF recommendation paper to define acceptance criteria for biomarker assays on the decisions taken from the assay data [18].Conclusion & future perspectiveWith this manuscript, the EBF wants to propose an open discussion whether it makes sense to move away from technology-based acceptance criteria in favor of decision-based acceptance criteria. We hope the discussion can get sufficient air time in industry, project teams and at upcoming meetings, either bioanalytically focused or with all stakeholders. We believe the proposal can alleviate the current ambiguity and nonadded value discussion on defining 'hybrid assay criteria'. Once integrated in our industry, harmonized decision-based acceptance criteria for bioanalytical assays in support of PK/safety will create a transparent platform to accept new technologies in the toolbox of the regulated bioanalytical (BA) scientist. And last but not least, the proposal should be seen as refining the criteria for studies 'in scope' of the guidelines. As advocated during the AAPS/EBF/JBF sister meetings [19,20], criteria of studies 'out of scope' should not automatically be held to these criteria but should be driven by scientific rationale considering decisions taken from the assay data.DisclaimerThe views and conclusion presented in this paper are those of the European Bioanalysis Forum and do not necessarily reflect the representative affiliation or company's position on the subject.Financial & competing interests disclosureThe authors have no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.No writing assistance was utilized in the production of this manuscript.References1 Shah VP, Midha KK, Dighe S et al. Analytical methods validation: bioavailability, bioequivalence and pharmacokinetic studies. Eur. J. Drug Metab. Pharmacokinet. 16(4), 249–255 (1991).Crossref, Medline, CAS, Google Scholar2 Shah VP, Midha KK, Findlay JW et al. Bioanalytical method validation – a revisit with a decade of progress. Pharm. Res. 17(12), 1551–1557 (2000).Crossref, Medline, CAS, Google Scholar3 Viswanathan CT, Bansal S, Booth B et al. Quantitative bioanalytical methods validation and implementation: best practices for chromatographic and ligand binding assays. Pharm. Res. 24(10), 1962–1973 (2007).Crossref, Medline, CAS, Google Scholar4 Fast DM, Kelley M, Viswanathan CT et al. Workshop report and follow-up – AAPS workshop on current topics in GLP bioanalysis: assay reproducibility for incurred samples – implications of Crystal City recommendations. AAPS J. 11(2), 238–241 (2009).Crossref, Medline, Google Scholar5 Booth B, Arnold ME, DeSilva B et al. Workshop report: Crystal City V – quantitative bioanalytical method validation and implementation: the 2013 revised FDA guidance. AAPS J. 17(2), 277–288 (2015).Crossref, Medline, CAS, Google Scholar6 Health Canada. Guidance for Industry: Conduct and Analysis of Bioavailability and Bioequivalence Studies – Part B: Oral Modified Release Formulations (1996). https://www.canada.ca/en/health-canada.html.Google Scholar7 US FDA. Guidance for Industry: Bioanalytical Method Validation (2018). https://www.fda.gov/downloads/drugs/guidances/ucm070107.Pdf.Google Scholar8 Japan Bioanalysis Forum. http://bioanalysisforum.jp/en.Google Scholar9 Timmerman P, Lowes S, Fast DM et al. Request for global harmonization of the guidance for bioanalytical method validation and sample analysis. Bioanalysis 2(4), 683 (2010).Link, CAS, Google Scholar10 Timmerman P, Arnold M, De Silva B et al. Introduction to the proposals from the Global Bioanalysis Consortium Harmonization Team. AAPS J. 16(6), 1159–1161 (2014).Crossref, Medline, Google Scholar11 ICH. Multidisciplinary guidelines. www.ich.org/products/guidelines/multidisciplinary/article/multidisciplinary-guidelines.Google Scholar12 Findlay JW, Smith WC, Lee JW et al. Validation of immunoassays for bioanalysis: a pharmaceutical industry perspective. J. Pharm. Biomed. Anal. 21(6), 1249–1273 (2000).Crossref, Medline, CAS, Google Scholar13 DeSilva B, Smith W, Weiner R et al. Recommendations for the bioanalytical method validation of ligand-binding assays to support pharmacokinetic assessments of macromolecules. Pharm. Res. 20(11), 1885–1900 (2003).Crossref, Medline, CAS, Google Scholar14 EBF meeting: large meets small. http://bru2011.europeanbioanalysisforum.eu/slides.Google Scholar15 EBF bioanalytical strategies for large molecules in modern drug development: LBA and LC–MS united. http://focus201706.europeanbioanalysisforum.eu/fw-slides/.Google Scholar16 Jenkins R, Duggan JX, Aubry AF et al. Recommendations for validation of LC–MS/MS bioanalytical methods for protein biotherapeutics. AAPS J. 17(1), 1–16 (2015).Crossref, Medline, CAS, Google Scholar17 Kringle RO. An assessment of the 4-6-20 rule for acceptance of analytical runs in bioavailability, bioequivalence, and pharmacokinetic studies. Pharm. Res. 11(4), 556–560 (1994).Crossref, Medline, CAS, Google Scholar18 Timmerman P, Herling C, Stoellner D et al. European Bioanalysis Forum recommendation on method establishment and bioanalysis of biomarkers in support of drug development. Bioanalysis 4(15), 1883–1894 (2012).Link, CAS, Google Scholar19 Industry input into ICH M10: experimental data as the cornerstone for a science driven bioanalytical guideline. http://focus201709.europeanbioanalysisforum.eu/slides/.Google Scholar20 Achieving a Harmonized Guideline for Bioanalytical Method Validation: ICH M10. AAPS News Magazine. https://www.aapsnewsmagazine.org/aapsnewsmagazine/articles/2018/may18/cover-story-may18.Google ScholarFiguresReferencesRelatedDetailsCited ByDirect bioanalysis or indirect calculation of target engagement and free drug exposure: do we apply double standards?Gregor Jordan & Roland F Staack10 February 2023 | Bioanalysis, Vol. 0, No. 0Feedback from the 8th European Bioanalysis Forum Young Scientist SymposiumLiesl Heughebaert, Adam Williams, Amelia Roberts, Cathy Jacobs, Claire Szuster, Connor Walker, Elien Van Nuffel, Esme Atkin, Francesca Minelli, Kate Groves, Katherine Sime, Katia Zeiser, Michele Protti, Lea Wagmann, Oscar Knight, Robert Stewart, Rosie Penford, Salvatore Calogero & Philip Timmerman3 February 2023 | Bioanalysis, Vol. 14, No. 23Biomarker context-of-use: how organizational design can impact the implementation of the appropriate biomarker assay strategyKyra J Cowan, Michaela Golob, Joanne Goodman, Anna Laurén, Lene Andersen, Philip De Decker, Lien Dejager, Marianne Scheel Fjording, Peter Groenen, Renaud Jasnowski, Nicole Justies, Matti Kimberg, Ulrich Kunz, James Lawrence, Mario Richter, Laetitia Sordé, Radboud van Trigt, Laurent Vermet, Alessandra Vitaliti, Michael Wright & Philip Timmerman29 July 2022 | Bioanalysis, Vol. 14, No. 13Recommendations and discussion points on immunogenicity, biomarkers, automation/technology and protein–MS from the 2021 European Bioanalysis Forum Focus WorkshopsPhilip Timmerman, Matthew Barfield, Kyra Cowan, Michaela Golob, Joanne Goodman, Ulrich Kunz, Anna Laurén, Iain Love, Robert Nelson, Roland F Staack, Johannes Stanta, Nico van de Merbel & Amanda Wilson4 October 2021 | Bioanalysis, Vol. 13, No. 19Supercritical fluid extraction–supercritical fluid chromatography of saliva: Single‐quadrupole mass spectrometry monitoring of caffeine for gastric emptying studies †16 August 2021 | Journal of Separation Science, Vol. 44, No. 19Bioanalytical method development and validation for establishing bioavailability and bioequivalenceUpdate to the European Bioanalysis Forum recommendation on biomarkers assays; bringing context of use into practiceJoanne Goodman, Kyra J Cowan, Michaela Golob, Lars Karlsson, Ulrich Kunz, Robert Nelson, Hans Ulrichts, Lauren Stevenson, Linda Terry & Philip Timmerman7 October 2020 | Bioanalysis, Vol. 12, No. 20European Bioanalysis Forum feedback on draft ICH M10 guideline on bioanalytical method validation during the Step 2b public consultation periodPhilip Timmerman, Joanne Goodman, Michaela Golob, Tobias Haslberger, Magnus Knutsson, Robert Nelson, Tom Verhaeghe & Steve White23 April 2020 | Bioanalysis, Vol. 12, No. 6sBioanalytical method validation: new FDA guidance vs. EMA guideline. Better or worse?Journal of Pharmaceutical and Biomedical Analysis, Vol. 165 Vol. 10, No. 16 Follow us on social media for the latest updates Metrics History Received 16 April 2018 Accepted 16 May 2018 Published online 21 August 2018 Published in print August 2018 Information© 2018 Newlands PressDisclaimerThe views and conclusion presented in this paper are those of the European Bioanalysis Forum and do not necessarily reflect the representative affiliation or company's position on the subject.Financial & competing interests disclosureThe authors have no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.No writing assistance was utilized in the production of this manuscript.PDF download

Circulating drug concentrations (clinical or preclinical) underly many interactions between industry and regulators; expressing safety coverage, pharmacokinetic-pharmacodynamic relationships or defining bioequivalence and dosing regimens. Accurate and precise measurement of these circulating concentrations is pivotal to the evolution and validation of any bioanalytical method that supports regulatory interactions. Since the bioanalyst is presented with a sub-aliquot of sampled biological matrix, how do they ensure this aliquot reflects the concentration in the subject at the time of collection? Here we share experiences from project support (internal and at CROs) that suggests we need to be ever vigilant translating the needs of bioanalysis with those of project teams. The simple mantra is for bioanalytical measurements to be physiologically relevant to the patient.

Tiered approach is rapidly gaining interest in the regulated bioanalytical community. Alternative approaches to the workflows as proposed in the regulatory Guidance (US FDA, EMA) are being used in discovery and early drug development, but with a growing array of assay types and studies requiring bioanalytical support in early drug development, the bioanalytical community is discussing how to bring best value to support these studies. Recently, international industry groups like European Bioanalysis Forum and Global Bioanalysis Consortium have discussed and published on the opportunity and need to include tiered approach more systematically in the early drug development support. On the back of these discussions, the Delaware Valley Drug Metabolism Discussion Group together with the European Bioanalysis Forum organized a meeting in Langhorne (PA, USA) to discuss the hurdles and added value of tiered approach with stakeholders from the Bioanalysis, quality assurance and PK community. The discussions focused on proposing scientific validation for studies where there is currently a mixed use of regulatory and tiered approach workflows. The meeting was well attended and the presentations and panel discussions contributed to a better understanding of what the industry is proposing as future practice.

In this article, we give feedback on the progress in industry on their efforts to provide practical and tangible solutions for a harmonized implementation of the principles of tiered approach. By describing tiered approach as different levels of scientific validation applied as an alternative to apply established regulatory validation principles [1–4] for studies where the guidance was not the intended scope, we hope to provide an acceptable handle for its adoption for an array of study types in industry. The principles of tiered approach became gradually known in regulated bioanalysis about a decade ago [5,6], received positive comments by Health Authority (HA) representatives [7], and were further propagated in regulated bioanalysis with the support of industry consortia [8,9]. Going forward, the bioanalytical community identified more areas in scope of application for tiered approach in a recently published special focus issue of Bioanalysis [10]. The practice of adopting less rigorous (or exhaustive) validation requirements in earlier stages of development, as highlighted in recent (draft) guidance [11] and confirmed during discussions at the recent Crystal City V meeting (Baltimore, MD, USA, 3–5 December 2013) stimulated the European Bioanalysis Forum (EBF) to organize a special workshop on Tiered Approach in June 2014 (Brussels, Belgium) to seek further alignment and provide practical solutions to bring tiered approach to the next level; in other words, in day-today practice. The aim of the workshop, which started from a simple paradigm that a validated assay does not necessarily equate to valid data, was fourfold:

BioanalysisVol. 6, No. 14 CommentaryEuropean Bioanalysis Forum continued plans to support liquid microsamplingPhilip Timmerman, Steve White, Zoe Cobb, Karen Woods, Ronald de Vries, Neil Spooner, Timothy Sangster, Lieve Dillen & Glen HawthornePhilip Timmerman*Author for correspondence: E-mail Address: ptimmerm@its.jnj.com Janssen Research & Development, Turnhoutseweg, 30, B2340 Beerse, Belgium, Steve White GlaxoSmithKline, Stevenage, UK, Zoe Cobb LGC, Fordham, UK, Karen Woods AstraZeneca, Alderly Park, Macclesfield, UK, Ronald de Vries Janssen Research & Development, Turnhoutseweg, 30, B2340 Beerse, Belgium, Neil Spooner GlaxoSmithKline, Stevenage, UK, Timothy Sangster Charles River laboratories, Edinburgh, UK, Lieve Dillen Janssen Research & Development, Turnhoutseweg, 30, B2340 Beerse, Belgium & Glen Hawthorne LGC, Fordham, UKPublished Online:27 Aug 2014https://doi.org/10.4155/bio.14.99AboutSectionsView ArticleView Full TextPDF/EPUB ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareShare onFacebookTwitterLinkedInRedditEmail View articleKeywords: consortiumEBFexperimentsliquid microsamplingvalidationReferences1 Abbott R, Smeraglia J, White S et al. Connecting strategies on dried blood spots. Bioanalysis 2(11), 1809–1816 (2010).Link, CAS, Google Scholar2 Timmerman P, White S, Globig S et al. EBF recommendation on the validation of bioanalytical methods for dried blood spots. Bioanalysis 3(14), 1567–1575 (2011).Link, CAS, Google Scholar3 Timmerman P, White S, Cobb Z et al. Update of the EBF recommendation for the use of DBS in regulated bioanalysis integrating the conclusions from the EBF DBS-microsampling consortium. Bioanalysis 5(17), 2129–2136 (2013).Link, CAS, Google Scholar4 Cobb Z, Ronald de Vries R, Spooner N et al. In-depth study of homogeneity in DBS using two different techniques: results from the EBF DBS-microsampling consortium. Bioanalysis 5(17), 2161–2169 (2013).Link, CAS, Google Scholar5 van Baar B, Verhaeghe T, Heudi O et al. IS addition in bioanalysis of DBS: results from the EBF DBS-microsampling consortium. 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Bioanalysis 6(5), 599–604 (2014).Link, CAS, Google ScholarFiguresReferencesRelatedDetailsCited ByDried Blood Spot in Laboratory: Directions and Prospects23 April 2020 | Diagnostics, Vol. 10, No. 4Feedback from the European Bioanalysis Forum liquid microsampling consortium: capillary liquid microsampling and assessment of homogeneity of the resultant samplesZoe Cobb, Morten Rohde, Iain Love, Valerie Boutet, Katrin Schroeter, Glen Hawthorne, Lieve Dillen, Matthew Barfield, Abdullah Kandira, Marion Kranenborgh, Stephen White & Philip Timmerman11 April 2019 | Bioanalysis, Vol. 11, No. 6Feedback from the European Bioanalysis Forum liquid microsampling consortium: microsampling: assessing accuracy and precision of handheld pipettes and capillariesGlen Hawthorne, Lieve Dillen, Matthew Barfield, Zoe Cobb, Abdullah Kandira, Katrin Schroeter, Steve White, Natasha A Karp, Iain Love, Valerie Boutet, Morten Rohde, Marion Kranenborgh & Philip Timmerman11 April 2019 | Bioanalysis, Vol. 11, No. 6Capillary microsampling in nonclinical safety assessment: practical sampling and bioanalysis from a CRO perspectiveDavid Coleman, Graeme Smith, Rachel Lawrence, Deborah McManus, Sunetha Diaram & Joanna Edwards19 May 2017 | Bioanalysis, Vol. 9, No. 10Is there a role for microsampling in antibiotic pharmacokinetic studies?3 May 2016 | Expert Opinion on Drug Metabolism & Toxicology, Vol. 12, No. 6Reducing pre-clinical blood volumes for toxicokinetics: toxicologists, pathologists and bioanalysts uniteKathryn Chapman, Josephine Burnett, Marco Corvaro, David Mitchell, Sally Robinson, Timothy Sangster, Susan Sparrow, Neil Spooner & Amanda Wilson19 November 2014 | Bioanalysis, Vol. 6, No. 22EBF: reflection on bioanalytical assay requirements used to support liquid microsamplingStephen White, Glen Hawthorne, Lieve Dillen, Neil Spooner, Karen Woods, Timothy Sangster, Zoe Cobb & Philip Timmerman20 November 2014 | Bioanalysis, Vol. 6, No. 19 Vol. 6, No. 14 STAY CONNECTED Metrics Downloaded 327 times History Published online 27 August 2014 Published in print July 2014 Information© Future Science LtdKeywordsconsortiumEBFexperimentsliquid microsamplingvalidationDisclaimerThe views and conclusion presented in this paper are those of the European Bioanalysis Forum and do not necessarily reflect the representative affiliations' or companys' positions on the subject.AcknowledgementsThe authors wish to thank the member companies of the EBF LMS Consortium (AstraZeneca, Charles River Laboratories, Covance, GlaxoSmithKline, Janssen Research & Development Novartis, LGC, Lundbeck, PRA, Sanofi, Shire, TNO Triskelion and QPS) for their contribution to the workshop and review of the manuscript and all EBF members for their input in the surveys.Financial & competing interests disclosureThe authors have no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.No writing assistance was utilized in the production of this manuscript.PDF download

Background: The European Bioanalysis Forum dried blood spots (DBS)/microsampling consortium is reporting back from the experiments they performed on further documenting the potential hurdles of the DBS technology. This paper is focused on the impact of hematocrit changes on DBS analyses. Results: The hematocrit can have an effect on the size of the blood spot, on spot homogeneity and on extraction recovery in a compound-dependent manner. The extraction recovery can change upon aging in an hematocrit-dependent way. Different card materials can give different outcomes. Conclusions: The results from the conducted experiments show that the issues of DBS in regulated bioanalysis are real and that the technology will need improvements to be ready for use as a general tool for regulated bioanalysis.

BACKGROUND:At the start of their work, the European Bioanalysis Forum dried blood spots microsampling consortium did not form a dedicated team to investigate the spot homogeneity. However, two teams performed experiments that produced results relating to sample homogeneity.RESULTS:The data, which were produced via two different approaches (a radiolabeled and a nonradiolabeled approach), are highly complementary and demonstrate clear effects on sample inhomogeneity due to the substrate type, compound and hematocrit levels.CONCLUSION:The results demonstrate that sample inhomogeneity is a significant hurdle to the use of dried blood spots for regulated bioanalysis that should be investigated further in the method establishment phase if the whole spot is not sampled.